Elbashir, S.M. et al. (2001). Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate. EMBO J. 20: 6877-6888. The length, overhangs, and sequences of synthetic siRNA are tested for their effects on silencing efficacy and the position of the mRNA cleavage site.

Hamilton, A.J. and Baulcombe, D.C. (1999). A species of small antisense RNA in posttranscriptional gene silencing in plants. Science 286: 950-952. The first recognition that siRNA are produced in the cell and are associated with gene silencing.

Schwarz, D.S. et al. (2003). Asymmetry in the assembly of the RNAi enzyme complex. Cell 115: 199-208. The relative stability of each end of the siRNA duplex is shown to be critical in determining which strand is loaded into the RISC. For knockdown to occur, the antisense strand must be loaded into the RISC.

RISC

Hammond, S.M. et al. (2000). An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature 404: 293-296. Describes the biochemical isolation of the RISC and the identification of a small RNA component.

Hammond, S.M. et al. (2001). Argonaute2, a link between genetic and biochemical analyses of RNAi. Science 293: 1146-1150. A member of the extensive Argonaute family of proteins genetically linked to gene silencing is identified as a component of the RISC.

Ambros, V. (2001). Development. Dicing up RNAs. Science 293: 811-813. Reviews the role of Dicer in the RNAi and miRNA pathways.

Bernstein, E. et al. (2001). Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409: 363-366. Identifies Dicer as the RNaseIII responsible for generation of siRNA in Drosophila extracts.

shRNA Plasmid Vectors

Brummelkamp, T.R. et al. (2002). A system for stable expression of short interfering RNAs in mammalian cells. Science 296: 550-553. The human H1 promoter is used to express shRNA from a plasmid vector in mammalian cells.

Off-Target and Non-Specific Effects

Persengiev, S.P. et al. (2004). Nonspecific, concentration-dependent stimulation and repression of mammalian gene expression by small interfering RNAs (siRNAs). RNA 10: 12-18. A large number of genes are shown to be non-specifically stimulated and repressed by synthetic siRNA.